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Bipolar CD4-targeted dual-DARPin-55/57 lipid nanoparticle enables efficient CRISPR/Cas-mediated HIV-1 DNA excision and reactivation blockade in latent CD4 T cell lines - PubMed

4 hours ago
  • #CRISPR/Cas
  • #HIV-1
  • #Lipid nanoparticle
  • A bipolar CD4-targeted dual-DARPin-55/57 lipid nanoparticle (LNP) was developed for efficient CRISPR/Cas-mediated HIV-1 DNA excision and reactivation blockade in latent CD4 T cell lines.
  • The LNP employs a unique bipolar conjugation strategy to decorate dual CD4-targeted DARPins (55 and 57) for selective uptake by resting CD4 T cells.
  • The system co-delivers spCas9-GFP mRNA (Sp9m) and HIV-1-specific single-guide RNAs (sgRNAs) targeting LTR and Gag (LGsg) into HIV-1 latently infected CD4 T cells.
  • In HIV-1 latency models (J-Lat 10.6 and 2D10 cell lines), the dual-DARPin-LNP efficiently excised integrated HIV-1 proviral DNA and blocked reactivation upon stimulation with latency-reversing agents.
  • This modular, non-viral, receptor-guided delivery platform represents a step toward next-generation curative interventions against persistent HIV-1 infection.